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First published online 28 March 2006
doi: 10.1242/jcs.02867

Journal of Cell Science 119, 1622-1631 (2006)
Published by The Company of Biologists 2006
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Research Article

NMDA induces post-transcriptional regulation of {alpha}2-guanylyl-cyclase-subunit expression in cerebellar granule cells

Sandra Jurado1, Fernando Rodríguez-Pascual2, José Sánchez-Prieto1, Francisco M. Reimunde2, Santiago Lamas2 and Magdalena Torres1,*

1 Departamento de Bioquímica, Facultad de Veterinaria, Universidad Complutense, Madrid, E-28040 Spain
2 Centro de Investigaciones Biológicas (CIB), Consejo Superior de Investigaciones Científicas (CSIC), Ramiro de Maeztu 9, Madrid, E-28040 Spain

* Author for correspondence (e-mail: mitorres{at}vet.ucm.es)

Accepted 4 January 2006

Activation of N-methyl-D-aspartate (NMDA) glutamate receptors commonly affects gene expression in different neurons. We reported previously that chronic treatment of rat cerebellar granule cells with NMDA (24 hours) upregulates the expression of mRNA encoding the {alpha}2 subunit of the nitric-oxide-sensitive guanylyl cyclase. However, the molecular mechanisms involved in this process remained to be elucidated. Here, we have performed mRNA-decay experiments using the transcriptional inhibitor actinomycin D, providing evidence that the half-life of {alpha}2 mRNA is significantly prolonged in cells exposed to NMDA. The role of the 3' untranslated region of the {alpha}2 transcripts in NMDA-induced mRNA stabilisation was examined and an association between the RNA-binding proteins AUF1 and ELAV-like protein 1 (HuR/HuA), and endogenous {alpha}2 mRNA was demonstrated in vivo, as revealed by coimmunoprecipitation experiments with specific antibodies against AUF1 and HuR. Further studies indicated that stimulation of the NMDA receptor induces a downregulation in AUF1 levels stabilising the {alpha}2 mRNA transcripts. These events are triggered through a mechanism that depends on formation of nitric oxide, and on the subsequent activation of guanylyl cyclase and cGMP dependent protein kinases.

Key words: AUF1, Cerebellar granule cells, mRNA decay, NO-sensitive guanylyl cyclase, N-methyl-D-aspartate


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